Cidofovir (CDV) is an acyclic nucleoside phosphonate analog that shows broad spectrum anti-DNA virus activity. In this study, we have investigated the influence of cidofovir on the tumor xenografts derived from HeLa and SiHa cells on nude mice. The HeLa/SiHa xenografts in nude mice were established by inoculating cells subcutaneously. Administration of cidofovir by intratumoral injection led to significant tumor reduction. Enhanced protein levels of p53 and p-pRb within the tumor samples were observed. Immunohistology analysis of the tumor sections indicated decreased PCNA index and increased apoptosis index. Our study gives more evidence and explanation on in vivo inhibition effect of cidofovir on HPV genome-positive cervical carcinoma cell line xenografts.

NF-κB is a transcription factor that induces the expression of inflammatory cytokines and antiapoptotic proteins. Earlier we designed a new NF-κB inhibitor, (−)-DHMEQ, and showed that it had potent anticancer and anti-inflammatory activities in various animal models without any toxicity. In the present research, we studied whether (−)-DHMEQ could be efficiently taken by cultured cells and irreversibly inhibit NF-κB by short time application to cultured cells. Even after mouse monocytic leukaemia RAW264.7 cells had been washed free of (−)-DHMEQ, lipopolysaccharide (LPS)-induced activation of NF-κB in these cells was still inhibited. Moreover, topical application for 15 min was found to induce dormancy of the cells against LPS for 2–8 h. When it was topically added to RAW264.7 cells in which NF-κB was activated by LPS, the inhibition lasted at least for 2 h. NF-κB directly upregulates expression of iNOS that produces NO. Short time application of (−)-DHMEQ also inhibited the function of cells in terms of NO production and iNOS induction in RAW264.7 cells. Thus, the fast incorporation of (−)-DHMEQ into the cells and irreversible inhibition of NF-κB by it were demonstrated, and this observation would explain its effective inhibition of certain functions in cellular and animal disease models.

In this study, the objective was to evaluate the presence of estrogen receptors α and β (ERα and ERβ) in cases of papillary carcinoma of the thyroid gland and to assess the practicality of this test. Immunohistochemical stains were performed for both ERα and ERβ, for evaluation of immunoreactivity in 90 papillary carcinomas. Three variables were evaluated in each sample: the intensity of the staining both nuclear and cytoplasmatic, and the spread of the stain over the sample. None of the histological samples showed immunoreactivity for ERα. Positive immunoreactivity results for ERβ were found in tissue samples in 66.6% (60 cases). The study shows that ERβ has no significant specification for differentiation between papillary carcinoma and other malignant lesions of the thyroid, while ERα is undetectable in this lesion. The ER testing in cases of papillary carcinoma of the thyroid gland is nonspecific and might be not necessary.

*First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui, Japan

The expression of survivin molecules has been confirmed in many types of cancer cells, including colon cancer cells, and they are considered important antiapoptotic molecules. Recent studies have revealed the existence of different splicing forms of survivin molecules; however, no studies have examined their expression in gastrointestinal cancers. In 2004, we reported the existence of the survivin-3B gene, a novel splice variant of survivin. In this study, we investigated the relationship between human colon cancer and our recently cloned survivin-3B gene with a coding region of 594 bp. In the first examination, survivin-3B expression was analyzed by RT-PCR in human colon cancer and adjacent normal mucosal tissues. The associations of its expression status with clinicopathological parameters and the prognosis were also examined. Survivin-3B mRNA expression was observed in 37 (46.3%) of 80 primary colon cancers, but not in the adjacent normal colonic mucosal tissue. The rate of survivin-3B gene expression was significantly higher in colon cancer with serosal invasion. The 5-year survival rate of patients with survivin-3B gene-positive primary colon cancer was significantly poorer, at 48.7%, than that (75.4%) of survivin-3B gene-negative patients. In the second examination, after the introduction of the survivin-3B gene into cells of the colon cancer cell line DLD-1, 5-fluorouracil-induced changes in their invasive capacity was examined. The invasion-inhibitory effect of 5-fluorouracil on survivin-3B gene-transfected DLD-1 cells was significantly lower than their empty vector gene-transfected counterparts. We speculate that survivin-3B expression in colon cancer is an important factor involved in the invasive capacity of cancer cells in the presence of anticancer drug.

Platinum-based chemotherapeutic drugs trigger apoptosis. and deregulation of apoptotic pathways may contribute to chemoresistance. We investigated the role of major Fas and Bcl-2 family members as predictors of response to platinum-based chemotherapy in epithelial ovarian cancer (EOC). The expression of Fas, FasL, FAP-1, Bax, Bcl-2, and Bcl-XL was analyzed in 35 women with EOC at the transcript level by semiquantitative RT-PCR and at the protein level by immunohistochemistry. The apoptotic index was determined by TUNEL assay. The response to chemotherapy was documented and at the end of six cycles of chemotherapy. Based on their response, two groups were identified: primary chemosensitive (n = 20) and primary chemoresistant (n = 15). Further, after a follow-up of 12–46 months, two groups were identified: no evidence of disease (n = 10) and evidence of disease (n = 25). The primary chemoresistant tumors in comparison to the chemosensitive tumors had significantly lower levels of Fas transcript (p = 0.026), Bax transcript (p = 0.042) and Bcl-2 protein (p = 0.038) and higher levels of Bcl-XL (p = 0.040). The apoptotic index revealed a significant inverse correlation only with Bcl-XL protein levels (p = 0.003). Patients with evidence of disease at last follow-up in comparison to those with no evidence of disease showed lower Bax transcript (p = 0.012), Bcl-2 protein (p = 0.014) and lower apoptotic index (p = 0.005) and higher Bcl-XL protein levels (p = 0.023). In conclusion, Bcl-2 family members and apoptotic index are useful in prediction of response to chemotherapy in EOC. These initial observations need to be validated in large-scale studies.

*Department of General Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China†Department of Biliary and Pancreatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China‡Department of Oncology, Zhongnan Hospital, Medical College of Wuhan University, Wuhan, China

Overexpression of the melanoma differentiation associated gene-7 (MDA-7)/IL-24 in vitro generally results in the growth suppression and induction of apoptosis of diverse human tumor cells. In this study, we investigated the effects of overexpression of the MDA-7/IL-24 gene in human hepatocellular carcinoma (HCC) cells in vitro and in vivo. Adenovirus-mediated overexpression of MDA-7 facilitated the MDA-7/IL-24-induced apoptosis and G2/M arrest in HCC cells, but not in the normal liver cell line L02, and the effect was independent of the p53 status. Inhibition of metastasis and angiogenesis was correlated with decreasing expression of STAT3, P-STAT3, MMP-2, VEGF, and TGF-β genes, regulated by STAT3 in MHCCLM6 cells. We also showed that Ad.mda-7 combined with doxorubicin (ADM) had significantly enhanced antitumor and antimetastatic effects in vivo, accompanied by the downregulation of VEGF, MMP-2, and TGF-β genes and the upregulation of E-cadherin genes. These data suggested that MDA-7/IL-24 induces its selective antitumor properties in HCC cells by promoting apoptosis independent of p53 status, inhibiting subcutaneous tumor growth and metastasis, and increasing the effect of chemotherapeutic agents. MDA-7/IL-24 represents a new class of cancer suppressor genes that may be useful in the targeted therapy of HCC.

*Department of Applied Life Sciences, Nihon University Graduate School of Bioresource Sciences, Kanagawa, Japan†Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon University, Kanagawa, Japan

Of the compounds contained in or derived from garlic (Allium sativum L.), alk(en)yl sulfides are known to be responsible for most of the physiological or neutraceutical functions of garlic. We previously found that diallyl trisulfide (DATS) is a potent inhibitor of tubulin polymerization and cancer cell growth, and an effective stimulator of the hepatic detoxification system. Here, we synthesized nine trisulfides having different aliphatic side chains, and determined their log P, a parameter for lipophilicity of nonionized solutes, and inhibitory activities, IC50 (μM), toward cancer cell growth. The log P values of these trisulfides ranged from the lowest, 2.72, for dimethyl trisulfide, to the highest, 7.62, for dipentyl trisulfide. The relationship between the IC50 and log P of the nine trisulfides was parabolic in nature, in which dibutenyl- and dipropyl-compounds, determined to have a log P of approximately 5, were located at the minimum point of the parabola, indicating the maximum potency. The reason why DATS, having a log P of about 4, was excessively stronger than diethyl trisulfide, with a similar log P, is not fully understood; but perhaps it can be explained by a higher reactivity of the diallyl compound in nucleophilic substitution. The compounds with 3-carbon chains were stronger in terms of growth inhibition than the others; but weaker compounds, those with 4- or 5-carbon chains, showed higher activity if a double bond was introduced into them to reduce their log P to the effective range. In this study, we confirmed the superiority of trisulfides with 3-carbon chains [i.e., DATS and dipropyl trisulfide (DPTS)]. In addition, we observed for the first time that dibutenyl trisulfide, a compound not found in garlic, is one of the potent structures among alk(en)yl trisulfides.

*Department of Medical Oncology, Faculty of Medicine, University of Gaziantep, Gaziantep, Turkey†Department of Physiology, Faculty of Medicine, University of Gaziantep, Gaziantep, Turkey‡Department of Medical Biology and Genetics, Faculty of Medicine, University of Gaziantep, Gaziantep, Turkey§Department of Pharmacology, Faculty of Medicine, University of Gaziantep, Gaziantep, Turkey¶Department of Medical Oncology, Faculty of Medicine, Erciyes University, Kayseri, Turkey#Department of Medical Oncology, Faculty of Medicine, Gazi University, Ankara, Turkey

The objective of this study was to analyze the genotype distributions and allele frequencies for ROCK2 Thr431Asn and Arg83Lys polymorphisms among breast cancer patients. In this case–control study, 223 patients with breast cancer were recruited and divided into two groups according to metastases (n = 128) and without metastases (n = 95). Genomic DNA from the patients and the control cases (n = 150) was analyzed by real-time PCR using a Light-Cycler. Neither genotype distributions nor the allele frequencies for the Arg83Lys polymorphism showed a significant difference between the groups. Although no marked changes were observed with nonmetastatic group, a statistically significant association was found between the control and metastatic group for the Thr431Asn polymorphism. Although homozygous carriers of the Thr431Thr genotype were more frequent, heterozygous carriers of the Thr431Asn genotype were less frequent among the metastatic patients than among controls. There was also an increase in Thr431 allele (60.5% in patients vs. 51.7% in controls) and decrease in Asn431 allele frequencies (48.3% in control vs. 39.5% in metastatic patients) in metastatic groups (p = 0.036). Our results demonstrate that Thr431Asn polymorphism of the ROCK2 gene could be a risk factor for the metastases of the breast cancer, and may help in predicting the prognosis.

Recent studies have revealed that hScrib, the human homolog of Drosophila Scribble, is an apical-basal polarity determinant and an essential component of the adherens junction. In addition, hScrib has a critical role in the inhibition of cell proliferation through cell cycle progression. hScrib has been reported to be involved in the processes of many cancers, such as breast cancer, cervical cancer, colon carcinoma, etc.; however, the correlation between hScrib and endometrial cancer has not been identified. To address a possible role of hScrib in the development of endometrial cancer, we examined the localization and expression of hScrib in endometrial cancer. The present study demonstrated that decreased expression and changed localization of hScrib were associated with clinical stage, histopathological differentiation, and lymph node metastasis in endometrial cancer. hScrib might share an adherens junction with basolateral membrane partially by acting on E-cadherin in endometrial cancer. This evidence suggests that hScrib is involved in the development of endometrial cancer.

Metronomic chemotherapy (MCT) is a novel therapeutic strategy for cancer treatment endowed with an antiangiogenic effect. It refers to regular administration of low doses of cytotoxic drugs, with minimal or no drug-free breaks. Previously, we demonstrated the immunomodulating activity of a single low-dose of cyclophosphamide (Cy) and the antitumor effect of MCT with Cy on established rat lymphomas and sarcomas. Here, we examined whether the immune response is responsible for the antitumor effect of MCT with Cy on L-TACB lymphoma. Inbred e rats and nude mice were subcutaneously challenged with L-TACB. After 7 days, they were distributed into two experimental groups: 1) treated animals, which were injected IP with Cy (10 mg/kg body weight) three times per week, and 2) control animals, which received IP saline injections. Exponential growth and decay and tumor doubling time were calculated. Also, serum IL-10 levels were measured. One hundred percent of treated rats showed tumor regression versus 0% of control rats. The increase of tumor-induced IL-10 levels was reverted by the treatment with Cy. On the other hand, there were no tumor regressions, in treated or control nude mice. However, the tumor doubling times of treated nude mice were significantly higher than those of control mice, implying that other antitumor mechanism(s), independent of the adaptive immune response, might be taking place. Our present results indicate that modulation of the immune response would be involved in the antitumor effect of MCT with Cy, because the absence of the specific immune response impairs, at least in part, its therapeutic effect in a lymphoma tumor model.

The long-term quality of life of patients after hematopoietic stem cell transplantation (HSCT) represents a multidisciplinary problem. HSCT can induce damage of various organs and tissues—from minimal potentially progressive subclinical changes to life-threatening conditions. Endocrine complications are among the most common late effects observed in survivors after HSCT. The relative risk of these complications is likely to be influenced by the underlying disease, type of therapy, and age at HSCT. Understanding the pathogenetic mechanisms of late complications that can occur after HSCT provides a basis for optimal surveillance and early intervention. Further research is needed for improved risk stratification of patients who are at low and high risk of developing late toxicity. Through collaboration between pathophysiologists, clinicians, and patient associations we can enhance the implementation of prospective studies and set forth effective preventive programs for survivors after HSCT.